PDBsum entry 2lr2

De novo protein

PDB id: 2lr2

Contents

Protein chain

76 a.a.

PDB id:

2lr2

Name:

De novo protein

Title:

Designed igg and lanthanide binding probe for solution nmr, mri and luminescence microscopy

Structure:

Immunoglobulin g-binding protein a. Chain: a. Engineered: yes

Source:

Artificial gene. Organism_taxid: 32630. Expressed in: escherichia coli. Expression_system_taxid: 469008.

NMR struc:

2 models

Authors:

A.W.Barb,T.G.Ho,J.H.Prestegard

Key ref:

A.W.Barb et al. (2012). Lanthanide binding and IgG affinity construct: potential applications in solution NMR, MRI, and luminescence microscopy. Protein Sci, 21, 1456-1466. PubMed id: 22851279

Date:

21-Mar-12 Release date: 12-Sep-12

Protein chain

No UniProt id for this chain

Struc: 76 a.a.

Protein Sci 21:1456-1466 (2012)

PubMed id: 22851279

Lanthanide binding and IgG affinity construct: potential applications in solution NMR, MRI, and luminescence microscopy.

A.W.Barb, T.G.Ho, H.Flanagan-Steet, J.H.Prestegard.

ABSTRACT

Paramagnetic lanthanide ions when bound to proteins offer great potential for structural investigations that utilize solution nuclear magnetic resonance spectroscopy, magnetic resonance imaging, or optical microscopy. However, many proteins do not have native metal ion binding sites and engineering a chimeric protein to bind an ion while retaining affinity for a protein of interest represents a significant challenge. Here we report the characterization of an immunoglobulin G-binding protein redesigned to include a lanthanide binding motif in place of a loop between two helices (Z-L2LBT). It was shown to bind Tb(3+) with 130 nM affinity. Ions such as Dy(3+) , Yb(3+) , and Ce(3+) produce paramagnetic effects on NMR spectra and the utility of these effects is illustrated by their use in determining a structural model of the metal-complexed Z-L2LBT protein and a preliminary characterization of the dynamic distribution of IgG Fc glycan positions. Furthermore, this designed protein is demonstrated to be a novel IgG-binding reagent for magnetic resonance imaging (Z-L2LBT:Gd(3+) complex) and luminescence microscopy (Z-L2LBT: Tb(3+) complex).